Structural, Functional and Molecular Dynamics Examination of a de novo cloned Otopetrin-like Proton Channel in crayfish.

Proton channels play a crucial role in many biological functions, as they are responsible for the selective transport of protons across cell membranes. Recently, Otopetrins, a family of eukaryotic proton-selective ion channels, have attracted significant attention due to their diverse physiological roles. Despite the importance of Otopetrins, their structural and functional properties remain relatively unexplored. As a model organism, crayfish have been extensively studied to gain insights into the functioning of the nervous system. These studies cover a wide range of aspects, including the properties of individual neurons and behavioral science. However, studying the physiological systems of crayfish poses challenges for molecular research due to limited molecular sequence information available for these organisms. In the present work was identified an originally cloned mRNA, coding an Otopetrin like proton channel in the crayfish. The coded protein was modeled in silico and possible conduction mechanisms and pathways were revealed. A plasmid of the cloned mRNA was heterologously expressed in HEK293T cells. Functional experiments on transfected cells indicated that the expressed mRNA was coupled to proton conduction across the cell membrane.

De Novo Cloning and Functional Characterization of a Mechanosensitive Piezo-Like Ion Channel in the Crayfish.

BACKGROUND/AIMS: Mechanosensitive ion channels are the principal elements in the transduction of mechanical force to neural activity. To date, considerably fewer studies have been published about the molecular and structural properties of mechanosensitive channels. Piezo channels are the only ion channel family in eukaryotes which is selectively gated by the membrane tension. Piezo channels have been described in mammals and some other eukaryotes. However, not much information is available for the crustaceans. METHODS: Conventional cloning methods were used to clone the putative PIEZO channel mRNA in crayfish ganglia samples. HEK293T cells were transfected by the plasmid of the cloned gene for functional studies. The CDS of the mRNA translated into the protein sequence and three-dimensional structure of the channel has been calculated. RESULTS: An mRNA, 9378 bp, was firstly cloned from crayfish which codes a 2674 residues protein. The cloned sequence is similar to the piezo channel mRNAs reported in the other species. The sequence of the coded protein has been analyzed, and some functional domains have been identified. A three-dimensional structure of the coded protein was successfully calculated in reference to mouse piezo 1 channel protein data. A plasmid with a fluorescent protein indicator was synthesized for heterologous expression in HEK293T cells. The evoked calcium response to mechanical stimulation was not different from those observed in the control cells. However, the transfected cells were more sensitive to the gating modifier YODA-1. CONCLUSION: Based on the apparent similarity in sequence, structure and functional properties to other known piezo channels, it has been proposed that cloned mRNA may code a piezo-like ion channel in crayfish.

Effect of different sealer activation techniques on dentinal tubule penetration.

The aim of this study was to evaluate the influence of different activation techniques on dentin tubule penetration of root canal sealer. Seventy-five teeth with single canals were chemomechanically prepared. A calcium silicate-based sealer was stained with a fluorescent dye (rhodamine B), placed into the canals and activated according to the following groups: control (no activation), EDDY, EndoActivator, ultrasonic and XP-Endo Finisher. Then, the samples were obturated. The percentages of sealer penetration at various depth levels of root sections were measured with confocal laser scanning microscopy. XP-Endo Finisher presented the highest penetration at 50 µm (p < 0.05). XP-Endo Finisher showed similar penetration with EDDY at 100 and 200 µm (p > 0.05) while presented higher penetration than the other groups (p < 0.05). At 500 µm, XP-Endo Finisher presented higher penetration than EndoActivator (p < 0.05) while similar penetration with the other groups (p > 0.05). XP-Endo Finisher can be recommended for activation during sealer placement for better penetration into dentin tubules.

Development of multistage recombinant protein vaccine formulations against toxoplasmosis using a new chitosan and porin based adjuvant system.

Toxoplasmosis is a global health problem affecting both human and animal populations. The lack of effective treatment makes the development of a vaccine against toxoplasmosis one of the main goals in the management of this disease. In our study, vaccine formulations containing the multistage recombinant antigens, rBAG1 + rGRA1 were developed with a combined adjuvant system consisting of chitosan and Salmonella Typhi porins in micro (MicroAS) and nanoparticulate (NanoAS) forms. BALB/c mice were immunized intraperitoneally with vaccine formulations two times at three-week intervals. Three weeks after the second vaccination, mice were challenged with 7-8 live tissue cysts of the virulent T. gondii PRU strain by oral gavage. Higher cellular uptake by macrophages and enhanced cellular (IFN-γ and I-4 in stimulated spleen cells) and humoral (IgG, IgG1, IgG2a) responses were obtained with the adjuvanted formulation, higher with microsystem when compared to that of nanosystem. Microsystem was found to stimulate Th1-polarized immune responses, whereasnon-adjuvanted antigens stimulated Th2-polarized immune response. The highest survival rate and reduction in cysts numbers and T. gondii DNA were obtained with the adjuvanted antigens.Our study showed that adjuvanted multistage recombinant vaccine systems increase theimmune response with strong protection againstT. gondii, more profoundly in microparticulate form.

Effect of mesenchymal stem cells therapy in experimental kaolin induced syringomyelia model.

BACKGROUND: Syringomyelia is a pathological cavitation of the spinal cord. In this study, we examined whether a syrinx cavity would limit itself with axonal regeneration and stem cell activity in the cavity, and we evaluated subjects on a functional basis. METHODS: Groups were designated as kaolin, trauma, kaolin-trauma, and saline groups. Also divided out of the syringomyelia treated groups were those given human mesenchymal stem cells (hMSCs). All groups were evaluated with immunohistochemistry, electron microscopy, confocal microscopy and functionally. RESULTS: The kaolin-trauma group had a significant correction of BBB score with hMSCs therapy. The syrinx cavity measurements showed significant improvement in groups treated with hMSCs. The tissue surrounding the syrinx cavity, however, appeared to be better organized in groups treated with hMSCs. The process of repair and regeneration of damaged axons in the lesion were more improved in groups treated with hMSCs. Using confocal microscopy, fluorescence of hMSCs was observed in the central canal, in the ependymal tissue, and around the lesion. CONCLUSIONS: It was concluded that axonal repair accelerated in groups receiving stem cells, and thus, stem cells may be effective in recovery of neural tissue and myelin damage in syringomyelia.

Knockout of zebrafish desmin genes does not cause skeletal muscle degeneration but alters calcium flux.

Desmin is a muscle-specific intermediate filament protein that has fundamental role in muscle structure and force transmission. Whereas human desmin protein is encoded by a single gene, two desmin paralogs (desma and desmb) exist in zebrafish. Desma and desmb show differential spatiotemporal expression during zebrafish embryonic and larval development, being similarly expressed in skeletal muscle until hatching, after which expression of desmb shifts to gut smooth muscle. We generated knockout (KO) mutant lines carrying loss-of-function mutations for each gene by using CRISPR/Cas9. Mutants are viable and fertile, and lack obvious skeletal muscle, heart or intestinal defects. In contrast to morphants, knockout of each gene did not cause any overt muscular phenotype, but did alter calcium flux in myofibres. These results point to a possible compensation mechanism in these mutant lines generated by targeting nonsense mutations to the first coding exon.

Chitosan Channels Stuffed with Mesenchyme Originated Stem/Progenitor Cells for Renovate Axonal Regeneration in Complete Spinal Cord Transection.

AIM: To examine the implantation of chitosan channels stuffed with mesenchyme-originated stem/progenitor cells (MSPCs) derived from adult rats in a spinal cord transection model. The level of axonal regeneration, the effect of chitosan channels on the survival of MSPCs, and the functional recovery results were also evaluated. MATERIAL AND METHODS: Chitosan channels stuffed with MSPCs were implanted at the level of T8 in a transected rat spinal cord. MSPCs were harvested from the pelvic bone marrow of adult rats, and the MSPC?chitosan channel group was compared with three control groups. The axonal regeneration capacity, the effect of chitosan channels on the survival of MSPCs, and the functional recovery results were compared among four groups. The survival of MSPCs was evaluated using histopathological techniques and electron microscopy, axonal regeneration/germination was evaluated by confocal microscopy, and locomotor function was assessed for 4 weeks using the Basso, Beattie, and Bresnahan locomotor score. RESULTS: The MSPC-chitosan channel group exhibited enhanced survival of transplanted MSPCs compared with MSPCs transplanted directly into the lesion cavity, although no significant difference was detected in locomotor function between the treatment and control groups. The MSPC-chitosan channel group demonstrated thicker myelination of axons than the other groups. CONCLUSION: Chitosan channels promoted the survival of transplanted MSPCs and created a tissue bridge after complete spinal cord transection. They also induced axonal regeneration and germination. No significant improvement in functional recovery was found between the groups.

Development and in vitro evaluation of a new adjuvant system containing Salmonella Typhi porins and chitosan.

In order to improve the immunogenicity of the highly purified vaccine antigens, addition of an adjuvant to formulation, without affecting the safety of the vaccine, has been the key aim of the vaccine formulators. In recent years, adjuvants which are composed of a delivery system and immunopotentiators have been preferred to induce potent immune responses. In this study, we have combined Salmonella Typhi porins and chitosan to develop a new adjuvant system to enhance the immunogenicity of the highly purified antigens. Cationic gels, microparticle (1.69 ± 0.01 µm) and nanoparticles (337.7 ± 1.7 nm) based on chitosan were prepared with high loading efficiency of porins. Cellular uptake was examined by confocal laser scanning microscopy, and the macrophage activation was investigated by measuring the surface marker as well as the cytokine release in vitro in J774A.1 macrophage murine cells. Porins alone were not taken up by the macrophage cells whereas in combination with chitosan a significant uptake was obtained. Porins-chitosan combination systems were found to induce CD80, CD86 and MHC-II expressions at different levels by different formulations depending on the particle size. Similarly, TNF-α and IL-6 levels were found to increase with porins-chitosan combination. Our results demonstrated that combination of porins with chitosan as a particulate system exerts enhanced adjuvant effect, suggesting a promising adjuvant system for subunit vaccines with combined immunostimulating activity.

Dentinal Tubule Penetration and Retreatability of a Calcium Silicate-based Sealer Tested in Bulk or with Different Main Core Material.

INTRODUCTION: The aim of this study was 2-fold: to evaluate the penetration of a tricalcium silicate-based endodontic sealer (EndoSequence BC Sealer; Brasseler USA, Savannah, GA) into dentinal tubules without a core material (sealer) or with .02 or .04 tapered bioceramic gutta-percha points and to compare the time required to remove the root fillings . METHODS: Roots of extracted human mandibular incisors (N = 60) were prepared with 0.04 taper nickel-titanium rotary files to #35 and randomly assigned into 3 groups (n = 10/group) according to the obturation method used: 1. obturating with sealer only, 2. sealer + .02 point, and 3. sealer + .04 point. The sealer was labeled with rhodamine B for analyzing dentinal tubule penetration under a confocal laser scanning microscope. The remaining specimens (n = 30) were used to measure the time for removal of the root canal fillings with retreatment files. The data were analyzed using 1-way analysis of variance and post hoc Games-Howell tests for dentinal tubule penetration and the Kruskal-Wallis test for retreatment time. RESULTS: Significantly greater sealer penetration and sealer-penetrated area was achieved when the sealer was used with a .04 gutta point (P < .05), whereas there was no difference between the sealer and .02 gutta point groups (P > .05). All test groups showed a similar depth of sealer penetration (P > .05). Groups with the gutta-percha points required a similar time to remove root filling (P > .05), whereas the working length could not be achieved in the sealer group. CONCLUSIONS: The use of a matched-taper bioceramic gutta-percha point enhanced the dentinal tubule penetration of the tested tricalcium silicate-based sealer. The use of a core material in conjunction with sealer facilitates removal of the root filling to the working length.

Effect of ultrasonic activation on dentinal tubule penetration of calcium silicate-based cements.

This study investigated the dentinal tubule penetration of mineral trioxide aggregate (MTA), NeoMTA Plus and Biodentine placed by either manual condensation or ultrasonic activation in simulated open apex model. Standardized divergent open apex models were created using palatal roots of 60 human maxillary molars and divided into six groups according to the used cements and activation methods (n = 10): MTA-manual condensation, MTA-ultrasonic activation, NeoMTA Plus-manual condensation, NeoMTA Plus-ultrasonic activation, Biodentine-manual condensation, Biodentine-ultrasonic activation. For the measurement of penetration, the cements were mixed with 0.1% Rhodamin B and 6-mm apical portions of each root canal were obturated in an orthograde direction. The roots were embedded into acrylic blocks, and 1-mm-thick sections were obtained at 3 mm from the apex. Specimens were mounted onto glass slides and scanned under a confocal laser scanning microscope (CLSM) and stereomicroscope. Dentinal tubule penetration areas, depth and percentage were measured using LSM and ImageJ software. The data were analyzed using two-way analysis of variance (anova) with Bonferroni correction (α = 0.05). No correlation was found between stereomicroscope and CLSM analyses (p > .05). CLSM analysis showed no significant differences between MTA, NeoMTA Plus, and Biodentine groups when manual condensation was used (p > .05). Ultrasonic activation did not increase the tubular penetration of MTA, NeoMTA Plus or Biodentine as compared to manual condensation of each material (p > .05). MTA, NeoMTA Plus and Biodentine showed similar tubular penetration when manual condensation was used. Ultrasonic activation of these cements had no effect on tubular penetration of each material as compared to the manual condensation counterparts.

Potential role of pyrin, the protein mutated in familial Mediterranean fever, during inflammatory cell migration.

Familial Mediterranean fever (FMF), the most common of the systemic autoinflammatory disorders, is caused by mutations in the MEFV (Mediterranean Fever) gene, which encodes the protein pyrin. Neutrophils, one of the major components during inflammation, are the main cell type that expresses pyrin. In response to an inflammatory stimulus, neutrophils migration to their main active site. To date, several pyrin-interacting proteins have been demonstrated to co-localise with the cytoskeletal protein actin, which is important in the process of neutrophil migration and raises the question of whether pyrin plays a role in the actin cytoskeletal network during inflammatory cell migration. In this study, we examined the possible role of pyrin during inflammatory cell migration in neutrophils. We generated a cell migration assay with neutrophils and primary neutrophils from patients. We also knocked down pyrin expression using siRNA and then performed cell migration assay. We showed co-localisation of pyrin and F-actin at the leading edge during inflammatory cell migration. In pyrin knocked down cells, we identified a significant decrease in neutrophil migration. In addition, we demonstrated a dramatic increase in migration in the neutrophils of FMF patients compared with a healthy control group. These data together provide new insight into the cellular function of pyrin and demonstrate an important link between pyrin and polymerising actin in the process of inflammatory cell migration.

Cloning of a putative sodium/calcium exchanger gene in the crayfish.

Crayfish is a common model animal for different experimental purposes. However, the lack of information about the genetic properties of the animal limits its use in comparison to other model animals. In the present study, a putative crayfish sodium/calcium exchanger gene has firstly been cloned in ganglia cDNA samples by conducting a series of PCR experiments, where a set of degenerate and specific primers and RACE method were used. The complete sequence is 2955 bp, and the ORF is 2718 bp in length. Molecular properties of the calculated peptide were similar to the sodium/calcium exchangers reported in the other species. Analysis of the qPCR data indicated that the putative gene has the highest expression level in the ganglia. However, an apparently elevated level of expression is observed in highly active tissues like heart, muscle and intestine, while the least expression level was observed in the stomach samples. It was proposed that the cloned gene may code the sodium/calcium exchanger protein in the crayfish.

Recombinant amelogenin regulates the bioactivity of mouse cementoblasts in vitro.

Amelogenin (AMG) is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG (rhAMG) on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts (OCCM-30) were treated with different concentrations (0.1, 1, 10, 100, 1000, 10,000, 100,000 ng · mL-1) of recombinant human AMG (rhAMG) and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen type I (COL I), osteopontin (OPN), runt-related transcription factor 2 (Runx2), cementum attachment protein (CAP), and alkaline phosphatase (ALP) genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer. Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomal-associated membrane protein-1 (LAMP-1), which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100,000 ng · mL-1 increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG (100,000 ng · mL-1) upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100,000 ng · mL-1 remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10,000 and 100,000 ng · mL-1 rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue.

Evaluation of dentinal tubule penetration depth and push-out bond strength of AH 26, BioRoot RCS, and MTA Plus root canal sealers in presence or absence of smear layer.

Background. This study compared the effect of smear layer on the penetration depth and push-out bond strength of various root canal sealers. Methods. A total of 90 extracted human mandibular premolars were assigned into 2 groups: smear layer preserved and smear layer removed. Then the roots were further divided into 3 subgroups according to the sealer tested: AH 26, BioRoot RCS and MTA Plus. Obturation was performed with gutta-percha and the relevant sealer was mixed with 0.1% rhodamine B. Three 1-mm-thick slices were obtained from the mid-third area of each root. Two slices were selected for the push-out test and the remaining slice was used to calculate the dentinal tubule penetration depth and percentage. Results. The retention of MTA Plus and BioRoot RCS was higher than that of AH 26 when the smear layer was preserved (P<0.05). BioRoot RCS showed the lowest penetration depth when the smear layer was removed (P<0.05). Conclusion. Dentinal tubule penetration of root canal sealers had a limited effect on their adhesion to root canal wall.

Efficacy of different irrigant protocols and application systems on sealer penetration using a stepwise CLSM analysis.

This study compared several irrigation protocols and application systems for sealer penetration into dentinal tubules. Single-rooted-human teeth were divided into 5 experimental groups (n = 15) and a control group (n = 5), according to final irrigation protocols: standard needle irrigation (SNI); Vibringe; Vibringe + NaviTip FX (Vibringe NFX); Endo Spray (ES); and passive-ultrasonic-irrigation (PUI). Following obturation of the root canals, the percentage of the sealer penetration was measured at different depths using stepwise CLSM analysis. The sealer penetration in the experimental groups was significantly higher than the control group at all levels (p < .05). No significant differences were observed between Vibringe and SNI or Vibringe NFX, ES, and PUI at all depths (p > .05). The Vibringe NFX, ES, and PUI groups allowed deeper sealer penetration than SNI at 100, 250, and 500 µm levels (p < .05). The irrigant activation, the needle design, and the application form (syringe or spray) may impact the quality of the seal that is achieved with root canal filling.

Fast calcium transients translate the distribution and conduction of neural activity in different regions of a single sensory neuron.

In the present study, cytosolic calcium concentration changes were recorded in response to various forms of excitations, using the fluorescent calcium indicator dye OG-BAPTA1 together with the current or voltage clamp methods in stretch receptor neurons of crayfish. A single action potential evoked a rise in the resting calcium level in the axon and axonal hillock, whereas an impulse train or a large saturating current injection would be required to evoke an equivalent response in the dendrite region. Under voltage clamp conditions, amplitude differences between axon and dendrite responses vanished completely. The fast activation time and the modulation of the response by extracellular calcium concentration changes indicated that the evoked calcium transients might be mediated by calcium entry into the cytosol through a voltage-gated calcium channel. The decay of the responses was slow and sensitive to extracellular sodium and calcium concentrations as well as exposure to 1-10 mM NiCl2 and 10-500 µM lanthanum. Thus, a sodium calcium exchanger and a calcium ATPase might be responsible for calcium extrusion from the cytosol. Present results indicate that the calcium indicator OG-BAPTA1 might be an efficient but indirect way of monitoring regional membrane potential differences in a single neuron.

Effect of a Low Surface Tension Vehicle on the Dentinal Tubule Penetration of Calcium Hydroxide and Triple Antibiotic Paste.

INTRODUCTION: The purpose of this study was to evaluate dentinal tubule penetration (DTP) of calcium hydroxide (CH) and triple antibiotic paste (TAP) when performed with distilled water (DW) or a low surface tension liquid (ie, propylene glycol [PG]). METHODS: Root apices of 40 single-rooted premolars were removed to obtain 14-mm roots in length. Root canals were enlarged to simulate immature teeth. After smear layer removal, the roots were randomly divided into 4 groups (n = 10) according to the root canal medicaments and the vehicles used: group 1:TAP + DW, group 2: TAP + PG, group 3: CH + DW, and group 4:CH + PG. Root canal medicaments were labeled with 0.1% rhodamine and applied into the canals using a Lentulo spiral. Specimens were molded into acrylic blocks, and 1-mm-thick sections were obtained from the middle third of each root. Specimens were mounted onto glass slides and scanned under a confocal laser scanning microscope. DTP depth, percentage, and area were measured using imaging software. Kruskal-Wallis and Mann-Whitney U tests were used for statistical analysis. The level of significance was set at P < .05. RESULTS: No significant difference was found among the experimental groups in terms of both percentage and depth of DTP (P > .05). CH had a lower penetration area compared with TAP regardless of the vehicle used (P < .05). CONCLUSIONS: A low surface tension vehicle did not alter the penetration of CH and TAP.

Push-out bond strength and dentinal tubule penetration of different root canal sealers used with coated core materials.

OBJECTIVES: The aim of this study was to compare the push-out bond strength and dentinal tubule penetration of root canal sealers used with coated core materials and conventional gutta-percha. MATERIALS AND METHODS: A total of 72 single-rooted human mandibular incisors were instrumented with NiTi rotary files with irrigation of 2.5% NaOCl. The smear layer was removed with 17% ethylenediaminetetraacetic acid (EDTA). Specimens were assigned into four groups according to the obturation system: Group 1, EndoRez (Ultradent Product Inc.); Group 2, Activ GP (Brasseler); Group 3, SmartSeal (DFRP Ltd. Villa Farm); Group 4, AH 26 (Dentsply de Trey)/gutta-percha (GP). For push-out bond strength measurement, two horizontal slices were obtained from each specimen (n = 20). To compare dentinal tubule penetration, remaining 32 roots assigned to 4 groups as above were obturated with 0.1% Rhodamine B labeled sealers. One horizontal slice was obtained from the middle third of each specimen (n = 8) and scanned under confocal laser scanning electron microscope. Tubule penetration area, depth, and percentage were measured. Kruskall-Wallis test was used for statistical analysis. RESULTS: EndoRez showed significantly lower push-out bond strength than the others (p < 0.05). No significant difference was found amongst the groups in terms of percentage of sealer penetration. SmartSeal showed the least penetration than the others (p < 0.05). CONCLUSIONS: The bond strength and sealer penetration of resin-and glass ionomer-based sealers used with coated core was not superior to resin-based sealer used with conventional GP. Dentinal tubule penetration has limited effect on bond strength. The use of conventional GP with sealer seems to be sufficient in terms of push-out bond strength.

Cloning and molecular characterization of a putative voltage-gated sodium channel gene in the crayfish.

Voltage-gated sodium channel genes and associated proteins have been cloned and studied in many mammalian and invertebrate species. However, there is no data available about the sodium channel gene(s) in the crayfish, although the animal has frequently been used as a model to investigate various aspects of neural cellular and circuit function. In the present work, by using RNA extracts from crayfish abdominal ganglia samples, the complete open reading frame of a putative sodium channel gene has firstly been cloned and molecular properties of the associated peptide have been analyzed. The open reading frame of the gene has a length of 5793 bp that encodes for the synthesis of a peptide, with 1930 amino acids, that is 82% similar to the α-peptide of a sodium channel in a neighboring species, Cancer borealis. The transmembrane topology analysis of the crayfish peptide indicated a pattern of four folding domains with several transmembrane segments, as observed in other known voltage-gated sodium channels. Upon analysis of the obtained sequence, functional regions of the putative sodium channel responsible for the selectivity filter, inactivation gate, voltage sensor, and phosphorylation have been predicted. The expression level of the putative sodium channel gene, as defined by a qPCR method, was measured and found to be the highest in nervous tissue.

Pyrin-PSTPIP1 colocalises at the leading edge during cell migration.

A set of mutations in the MEditerranean FeVer (MEFV) gene causes familial Mediterranean fever (FMF), the most common auto-inflammatory disease. The gene encodes a protein named pyrin, which appears to play an important role in inflammatory pathways. Furthermore, pyrin, which is expressed in neutrophils, has been reported to interact with proline-serine-threonine phosphatase-interacting protein 1 (PSTPIP1) and actin proteins. However, the relations between pyrin and PSTPIP1 during the cell migration have not yet been elucidated. In the present study, we constructed a cell migration assay method using HL-60 cells. Pyrin-PSTPIP1 interactions were analysed by immunofluorescence staining in control, differentiated and differentiated-stimulated HL-60 cells. In stimulated cells, pyrin-polymerised actin, PSTPIP1-polymerised actin and pyrin-PSTPIP1 were found to be colocalised. Pyrin has been shown to be colocalised with actin and PSTPIP1 at the leading edge of the migrating cell. For the first time, PSTPIP1 was found to interact with dynamic actin and pyrin at the site of polarisation.